{"title":"人Survivin的构建、表达及其多克隆抗体的制备","authors":"Hai-hong ZHANG, Xi-zhen ZHANG, Dong-hai ZHAO, He-liang SHI, Yong-hui YU, Yong-ge WU, Xiang-hui YU, Wei KONG","doi":"10.1016/S1005-9040(09)60024-6","DOIUrl":null,"url":null,"abstract":"<div><p>Survivin, a novel member of inhibitor of apoptosis(IAP) protein family, is aberrantly expressed in cancer but undetectable in normal, differentiated adult tissues. The cancer-specific expression of survivin, coupled with its importance in inhibiting cell death and in regulating cell division makes it a useful diagnostic marker of cancer and a potential target for cancer treatment. Survivin cDNA amplified from the total RNA of 293 cells through RT-PCR was cloned into prokaryotic expression vector pRSET-B. The recombinant plasmid pRSET-B-Surv was expressed in <em>E.coli</em> BL21, and the relative molecule mass(<em>M</em><sub>r</sub>) of expressed fusion protein was approximately 21000. The recombinant protein was purified through Ni<sup>2+</sup> affinity chromatography column and characterized by SDS-PAGE and Western blot. The purified recombinant protein was then injected into rabbits, and antisurvivin polyclonal antibody with a high titer was obtained.</p></div>","PeriodicalId":9785,"journal":{"name":"Chemical Research in Chinese Universities","volume":"24 6","pages":"Pages 767-770"},"PeriodicalIF":3.1000,"publicationDate":"2008-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1005-9040(09)60024-6","citationCount":"2","resultStr":"{\"title\":\"Construction and Expression of Human Survivin and Preparation of Its Polyclonal Antibody\",\"authors\":\"Hai-hong ZHANG, Xi-zhen ZHANG, Dong-hai ZHAO, He-liang SHI, Yong-hui YU, Yong-ge WU, Xiang-hui YU, Wei KONG\",\"doi\":\"10.1016/S1005-9040(09)60024-6\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Survivin, a novel member of inhibitor of apoptosis(IAP) protein family, is aberrantly expressed in cancer but undetectable in normal, differentiated adult tissues. The cancer-specific expression of survivin, coupled with its importance in inhibiting cell death and in regulating cell division makes it a useful diagnostic marker of cancer and a potential target for cancer treatment. Survivin cDNA amplified from the total RNA of 293 cells through RT-PCR was cloned into prokaryotic expression vector pRSET-B. The recombinant plasmid pRSET-B-Surv was expressed in <em>E.coli</em> BL21, and the relative molecule mass(<em>M</em><sub>r</sub>) of expressed fusion protein was approximately 21000. The recombinant protein was purified through Ni<sup>2+</sup> affinity chromatography column and characterized by SDS-PAGE and Western blot. The purified recombinant protein was then injected into rabbits, and antisurvivin polyclonal antibody with a high titer was obtained.</p></div>\",\"PeriodicalId\":9785,\"journal\":{\"name\":\"Chemical Research in Chinese Universities\",\"volume\":\"24 6\",\"pages\":\"Pages 767-770\"},\"PeriodicalIF\":3.1000,\"publicationDate\":\"2008-11-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/S1005-9040(09)60024-6\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Chemical Research in Chinese Universities\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1005904009600246\",\"RegionNum\":4,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, MULTIDISCIPLINARY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chemical Research in Chinese Universities","FirstCategoryId":"92","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1005904009600246","RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
Construction and Expression of Human Survivin and Preparation of Its Polyclonal Antibody
Survivin, a novel member of inhibitor of apoptosis(IAP) protein family, is aberrantly expressed in cancer but undetectable in normal, differentiated adult tissues. The cancer-specific expression of survivin, coupled with its importance in inhibiting cell death and in regulating cell division makes it a useful diagnostic marker of cancer and a potential target for cancer treatment. Survivin cDNA amplified from the total RNA of 293 cells through RT-PCR was cloned into prokaryotic expression vector pRSET-B. The recombinant plasmid pRSET-B-Surv was expressed in E.coli BL21, and the relative molecule mass(Mr) of expressed fusion protein was approximately 21000. The recombinant protein was purified through Ni2+ affinity chromatography column and characterized by SDS-PAGE and Western blot. The purified recombinant protein was then injected into rabbits, and antisurvivin polyclonal antibody with a high titer was obtained.
期刊介绍：
The journal publishes research articles, letters/communications and reviews written by faculty members, researchers and postgraduates in universities, colleges and research institutes all over China and overseas. It reports the latest and most creative results of important fundamental research in all aspects of chemistry and of developments with significant consequences across subdisciplines.
Main research areas include (but are not limited to):
Organic chemistry (synthesis, characterization, and application);
Inorganic chemistry (bio-inorganic chemistry, inorganic material chemistry);
Analytical chemistry (especially chemometrics and the application of instrumental analysis and spectroscopy);
Physical chemistry (mechanisms, catalysis, thermodynamics and dynamics);
Polymer chemistry and polymer physics (mechanisms, material, catalysis, thermodynamics and dynamics);
Quantum chemistry (quantum mechanical theory, quantum partition function, quantum statistical mechanics);
Biochemistry;
Biochemical engineering;
Medicinal chemistry;
Nanoscience (nanochemistry, nanomaterials).
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